5J7A

Lipidic cubic phase injector is a viable crystal delivery system for time-resolved serial crystallography.

Nogly, P., Panneels, V., Nelson, G., Gati, C., Kimura, T., Milne, C., Milathianaki, D., Kubo, M., Wu, W., Conrad, C., Coe, J., Bean, R., Zhao, Y., Bath, P., Dods, R., Harimoorthy, R., Beyerlein, K.R., Rheinberger, J., James, D., DePonte, D., Li, C., Sala, L., Williams, G.J., Hunter, M.S., Koglin, J.E., Berntsen, P., Nango, E., Iwata, S., Chapman, H.N., Fromme, P., Frank, M., Abela, R., Boutet, S., Barty, A., White, T.A., Weierstall, U., Spence, J., Neutze, R., Schertler, G., Standfuss, J.

(2016) Nat Commun 7: 12314-12314

• PubMed: 27545823 Search on PubMedSearch on PubMed Central
• DOI: https://doi.org/10.1038/ncomms12314
• Primary Citation of Related Structures:  
  5J7A


• PubMed Abstract: 
  

  Serial femtosecond crystallography (SFX) using X-ray free-electron laser sources is an emerging method with considerable potential for time-resolved pump-probe experiments. Here we present a lipidic cubic phase SFX structure of the light-driven proton pump bacteriorhodopsin (bR) to 2.3?? resolution and a method to investigate protein dynamics with modest sample requirement. Time-resolved SFX (TR-SFX) with a pump-probe delay of 1?ms yields difference Fourier maps compatible with the dark to M state transition of bR. Importantly, the method is very sample efficient and reduces sample consumption to about 1?mg per collected time point. Accumulation of M intermediate within the crystal lattice is confirmed by time-resolved visible absorption spectroscopy. This study provides an important step towards characterizing the complete photocycle dynamics of retinal proteins and demonstrates the feasibility of a sample efficient viscous medium jet for TR-SFX.

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Organizational Affiliation: 

Laboratory for Biomolecular Research, Paul Scherrer Institute, Villigen 5232, Switzerland.